Abstract
Aim:
To establish a steady purification method for producing recombinant humanized anti-HBsAg Fab from yeast fermentation supernatant.
Methods:
Humanized anti-HBs scFv was used to immunize BALB/c mice to obtain anti-scFv monoclonal antibody (mAb) which was used to purify recombinant humanized anti-HBsAg Fab from yeast fermentation supernatant by affinity chromatography.
Results:
The purity of purified recombinant anti-HBsAg Fab was above 95% and its recovery rate was about 75%-85%.
Conclusion:
An efficient affinity chromatography suitable for purification of recombinant humanized anti-HBsAg Fab in large-scale was established.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibody Affinity
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Chromatography, Affinity
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Fermentation
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Hepatitis B Antibodies / immunology
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Hepatitis B Antibodies / isolation & purification*
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Hepatitis B Surface Antigens / immunology*
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Hybridomas / metabolism
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Immunization
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Immunoglobulin Fab Fragments / immunology
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Immunoglobulin Fab Fragments / isolation & purification*
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Immunoglobulin Fragments / immunology
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Immunoglobulin Fragments / isolation & purification*
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Immunoglobulin Variable Region / immunology
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Immunoglobulin Variable Region / isolation & purification
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Mice
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Mice, Inbred BALB C
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Pichia / immunology
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Recombinant Proteins / immunology
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Recombinant Proteins / isolation & purification*
Substances
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Antibodies, Monoclonal
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Hepatitis B Antibodies
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Hepatitis B Surface Antigens
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Immunoglobulin Fab Fragments
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Immunoglobulin Fragments
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Immunoglobulin Variable Region
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Recombinant Proteins
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immunoglobulin Fv